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ARTICLE NO. RC976622
Sanetaka Shirahata, Shigeru Kabayama, Mariko Nakano, Takuni Miura,
Kenichi Kusumoto, Miho Gotoh,
Hidemitsu Hayashi,* Kazumichi Otsubo,**
Institute of Cellular Regulation Technology, Graduate School of Genetic Resources Technology, Kyushu University, 6-10-1
Hakozaki, Higashi-ku, Fukuoka 812-81, Japan; *Water Institute, Nisshin Building 9F, 2-5-10 Shinjuku, Tokyo 160,
Japan; and **Nihon Trim Co. Ltd., Meiji Seimei Jusou Building 6F, 1-2-13 Shinkitano, Yodogawa-ku, Osaka 532, Japan
Active oxygen species or free radicals are considered to cause extensive oxidative damage to biological macromolecules, which brings about a variety of diseases as well as aging. The ideal scavenger for active oxygen should be ‘active hydrogen’. ‘Active hydrogen’ can be produced in reduced water near the cathode during electrolysis of water. Reduced water exhibits high pH, low dissolved oxygen (DO), extremely high dissolved molecular hydrogen (DH), and extremely negative redox potential (RP) values. Strongly electrolyzed-reduced water, as well as ascorbic acid, (+)-catechin and tannic acid, completely scavenged O2•- produced by the hypoxanthine-xanthine oxidase (HX-XOD) system in sodium phosphate buffer (pH 7.0).
The superoxide dismutase (SOD)-like activity of reduced water is stable at 4°C for over a month and was not lost even after neutralization, repeated freezing and melting, deflation with sonication, vigorous mixing, boiling, repeated filtration, or closed autoclaving, but was lost by opened autoclaving or by closed autoclaving in the presence of tungsten trioxide which efficiently adsorbs active atomic hydrogen. Water bubbled with hydrogen gas exhibited low DO, extremely high DH and extremely low RP values, as does reduced water, but it has no SOD-like activity. These results suggest that the SOD-like activity of reduced water is not due to the dissolved molecular hydrogen but due to the dissolved atomic hydrogen (active hydrogen). Although SOD accumulated H2O2 when added to the HX-XOD system, reduced water decreased the amount of H2O2 produced by XOD. Reduced water, as well as catalase and ascorbic acid, could directly scavenge H2O2. Reduced water suppresses single-strand breakage of DNA by active oxygen species produced by the Cu(II)-catalyzed oxidation of ascorbic acid in a dose-dependent manner, suggesting that reduced water can scavenge not only O2•- and H2O2, but also 1O2 and •OH.
Abbreviations: AET, 2-(aminoethyl)isothiuroniumAsA, ascorbic acid; CL, chemiluminescence; CLA, Cypridinaluciferin analog; DO, dissolved oxygen; DH, dissolved hydrogen, EC, electrical conductance; HX, hypoxanthine; RP, redox potential; SOD, superoxide dismutase; XOD, xanthine oxidase;